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The effect of pyrazinederivatives on secondarymetabolitesproductionin in vitro cultures
Klepalová, Libuše ; Tůmová, Lenka (advisor) ; Kovalíková, Zuzana (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate Mgr. Libuše Klepalová Consultant Doc. PharmDr. Lenka Tůmová CSc. Title of Thesis The effect of pyrazine derivatives on secondary metabolites production in in vitro cultures This work studies the possibility of influencing the production of silymarin and taxifoline in in vitro cultures of Silybum marianum L. using the elicitation method. The pyrazine derivate BBPS3 in three different concentrations (c1-3.3529·10-3 mol/l, c2-3.3529 ·10-4 mol/l and c3-3.3529·10-5 mol/l) was used as elicitor. Sampling took place at 6, 24, 48, 72 and 168 hours after application of the substance. Control samples were left without the use of an elicitor. At the same time, samples of the nutrient medium were taken. The analysis was performed using the HPLC method. Thus, in order to increase the content of silymarin complex and taxifoline, it seemed more appropriate to use a callus culture, compared to a suspension culture. The statistically most significant increase in the content of silymarin complex in callus culture was achieved after 72 h of elicitor treatment with a concentration of c2 (80.7 µg/g DW). Statistically the most significant increase in the taxifoline content occurred after 168 h of elicitation of the...
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Analytical and bioanalytical study of selected drugs using UHPLC
Klepalová, Libuše ; Kovaříková, Petra (advisor) ; Kučera, Radim (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of pharmaceutical chemistry and drug analysis Candidate: Libuše Klepalová Supervisor: doc. PharmDr. Petra Kovaříková, Ph.D. Title of the Master's Thesis: Analytical and bioanalytical study of selected drugs using UHPLC Abstract High-performance liquid chromatography coupled with mass spectrometry plays a crucial role in drug analysis from biological materials, mainly due to its sensitivity, specificity and ability to provide structural information on the analytes. Multiple myeloma is a serious hematologic disease, accounting for about 1 % of all oncological diseases. New drugs from the group of proteasome inhibitors namely, carfilzomib and bortezomib represent a great promises for patients with relapsing multiple myeloma or in the case of the resistant to other anticancer drugs. The aim of this work was to develop pilot chromatographic conditions for HPLC-MS analysis of carfilzomib and bortezomib and evaluate different extraction of these drugs from plasma. Separation was carried out on an Ascentis column C18 10 x 3 mm, 3 µm, (Supelco, Germany). Electrospray ionization with ion trap mass analyzer were used in both cases. The mobile phase consisting of water and acetonitrile (20:80) was the most appropriate for carfilzomib. The flow rate of...
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